The goal of this Phase I SBIR proposal is to test the feasibility of an inherently broad-spectrum antibacterial tissue scaffolding matrix in preventing and eliminating biofilms while improving wound closure in chronic wounds. Delayed non-healing chronic wounds are responsible for approximately 100,000 annual non-traumatic lower-limb amputations occurring in the United States alone. Nearly 70% of all chronic wounds contain microbial biofilms which further complicate wound care by delaying healing and releasing inflammatory cytokines. In addition, the emergence of multi drug resistant organisms (MDROs) have made the current topical and systemic antibiotics progressively less effective in combating chronic wounds. However, the biological scaffolds and matrices promote wound healing, but are ineffective at eliminating pathogens thereby complicating wound care. Thus, there are no current products to promote wound healing while eliminating and preventing biofilms. G4F-1, our proposed product can (i) prevent/eliminate biofilms through a unique mechanism of action that is broad spectrum antibacterial, and (ii) promote tissue regeneration by providing cell attachment sites within the scaffolding matrix. Unlike any of the products currently used in the clinics, G4F-1 does not rely on using any externally added antibiotics, but takes advantage of charge composition to exert toxicity against Gram - positive and -negative bacteria (including antibiotic resistant strains). Furthermore, G4F-1 relies only on sequence design for antimicrobial charge ratio, and therefore is cell friendly, non-inflammatory and nontoxic with a structure and pore size very similar to the native extracellular matrix (ECM). Hence, in this feasibility study we propose three specific aims: Specific Aim1. Evaluate the efficacy of two different active formulations of PEP class gels against established Staphylococcus aureus and Pseudomonas aeruginosa biofilms in vitro. Specific Aim2. Demonstrate Staphylococcus aureus and Pseudomonas aeruginosa bacterial and biofilm prevention/reduction ex vivo. Specific Aim3. Demonstrate Staphylococcus aureus and Pseudomonas aeruginosa bacterial and biofilm reduction in vivo using a chronic wound porcine model. It is expected that the generated data will provide the necessary information for a phase II study to test polymicrobial biofilms, validate the product in larger statistically significant group of animals and enable commercialization of the proposed product. Phase II studies will also enable GMP manufacturing and execute GLP studies in preparation for FDA submission.